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Enter a sequence to perform a BLAST-based search of Addgene plasmid sequence databases.

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We narrowed to 1,045 results for: ins;

Showing: 431 - 440 of 1045 results

  1. Addgene’s Viral Service - Why Virus? Why Now?

    Type
    Blog Post
    ...Root from the Broad Institute and Alex Chavez from the Wyss Institute were instrumental in helping us establish...School, and Jonathan Ting’s lab from the Allen Institute have supported the AAV initiative from the beginning...Janelia Research Campus have provided valuable insight into AAV production. Finally, we are blessed to...permission from relevant plasmid depositors and their institutions to distribute viral preps derived from plasmids...

  2. Plasmids 101: Sequence and Ligation Independent Cloning (SLIC)

    Type
    Blog Post
    ...restriction enzyme cloning uses short sticky ends to insert genes into plasmid backbones, base pairing between...absence of the other dNTPs. Once the vector and insert are (separately) digested to create 10–12 base ...on the power of homologous recombination. SLIC inserts and linearized vectors must have homologous DNA...imprecise T4 digestion (no dNTPs needed), allows both insert and vector to be digested in the same tube. Alternatively...these approaches require higher concentrations of insert to succeed. 5′ overhangs are generated in the linearized... be used with different types of PCR-generated inserts. By harnessing the power of DNA repair, you can...

  3. Viral Vectors 101: Viral Vector Elements

    Type
    Blog Post
    ...interact with the host. This is the role of Gag proteins: they contain the matrix, capsid, and nucleocapsid...assembly, and display. Pol, the last component, contains the reverse transcriptase and integrase components...convert the cargo from RNA to DNA and permanently insert the cargo into the host genome. Lentiviral-specific... Viral packaging and entry The viral packaging proteins for AAV are rep and cap. Rep is required for viral...discussed. LXs/Iva2 are primarily capsid assembly proteins required for packaging of the virus into particles...Resources Kulanayake, S., Tikoo, S. K. Adenovirus Core Proteins: Structure and Function. Viruses. 2021 13(3):388...

  4. Viral Vectors 101: Types of viruses

    Type
    Blog Post
    ...proto-oncogenes and disrupt their expression (Robinson, 1982). To add insult to injury, retroviruses generally have...This viral DNA (also called proviral DNA) is then inserted into the host's genome and is used to make more...virus-specific elements such as the LTRs or structural proteins are not interchangeable between the two systems...Adenovirus generally has high immunogenicity, but some strains have been engineered to induce a lower inflammatory...immunogenicity low! References and Resources References Robinson HL. Retroviruses and cancer. Rev Infect Dis. 1982...

  5. Plasmids 101: Methylation and Restriction Enzymes

    Type
    Blog Post
    ...if not methylated (if it comes from a phage for instance). Aside from restriction modification systems,...effectively cleave DNA. Common lab E. coli K12 strains such as DH5alpha contain 3 methylases that recognize...Dam or Dcm methylation, you can ensure this site remains unblocked by first cloning your DNA into a dam–...and re-purifying it. These specialized E. coli strains have been specifically engineered to be Dam and...those sites. Please keep in mind that dam-/dcm- strains may have an increased rate of mutation (as these...in mis-match repair functions of Dam), so these strains should not be used for long term storage. Additional...

  6. How to Keep a Lab Notebook for Bioinformatic Analyses

    Type
    Blog Post
    ...another name for a folder. For instance, your Desktop directory contains the folders and files that you...including background on other analyses that have inspired you to perform this analysis. Having a clearly...multiple projects in a given day. For example, instead of naming a file “output” name the file “1_output...subsequent word in a file name after the first word begins with a capital letter and words are not separated...temporarily used. Just be sure that the original code remains in tact! Take notes Comment in your scripts: ...

  7. Protein Tagging with CRISPR/Cas9: A Conversation with Mendenhall and Myers

    Type
    Blog Post
    ...small percentage of the proteins inside a cell have a commercial antibody against them. There’s also a question...Mendenhall of the University of Alabama in Huntsville explains it, a major goal in his laboratory is to understand...track down every last one of those gene regulatory proteins and the places in the genome where they can be.... If things work correctly, the second plasmid inserts a small epitope tag called FLAG at the end of the...

  8. Pooled CRISPR Libraries Offer Genome-Wide Control for Large-Scale Functional Screens

    Type
    Blog Post
    ...and control genes, but according to the Broad Institute's Silvana Konermann, the first generation of CRISPR-Cas9...recognized as an ideal spot to fuse the activators instead of at the protein’s C terminus. With that new design... and knew she was on the right track. The next insight was to assemble a synthetic transcription activation...complex consisting of multiple distinct effector domains that work together synergistically, modeled after...CRISPRa) sgRNA library uses the sunCas9 system and contains 10 sgRNAs for each transcription start site in...The transcriptional repressor (CRISPRi) library contains 10 sgRNAs for each transcription start site in...

  9. Plasmids 101: Yeast Vectors

    Type
    Blog Post
    ...occurring resistant mutants and the insensitivity of yeast strains to some antibiotics. Auxotrophy is ...bacteria, yeast can post-translationally modify proteins yet they still share many of the same technical...replication (Please note: This first section primarily pertains to ORIs in budding yeast, Saccharomyces cerevisiae...other hand, do not require a well defined ORI. Instead, the size and A-T content of the DNA (apparently...compound required for its growth. Many auxotrophic strains of yeast exist which can be easily maintained when...

  10. Plasmids 101: Modular Cloning Applications and Kits

    Type
    Blog Post
    ...multiplexing of sgRNAs against unique genomic targets or multiple sgRNAs against the same target to ensure...protein purification and metabolic engineering. If instead you want to build genetic circuits and would benefit...library of parts but don’t need to add tags to your proteins, then the CIDAR Kit is an excellent option (Iverson...system includes a toolkit to express recombinant proteins within the mitochondrial matrix.     Figure...can be repurposed for CRISPR applications (gene insert = Cas9 + gRNA), Addgene distributes specialized...of a Toolkit for the Expression of Heterologous Proteins in Yeast Mitochondria.” BMC Biotechnology, vol...
Showing: 431 - 440 of 1045 results